Re: [Histonet] Re: Superfrost slides-Histonet question

From:

I.B.

Mari Ann,
Yes, quite soft tissue. Freezing took a while to figure out, but I now do
it in much the same manner as the Paper Embedding technique at Pathology
Innovations. The difficulty is in acting quickly enough for proper contact
and freezing. I have tried freezing the tissue in 200 proof ethanol in Li
N2, but apparently I am doing something wrong, as the tissue and OCT
compound around it simply refuse to cut (even though it is allowed to come
to cabinet temperature for eight hours before I attempt to section it). I
am sure the error is on my end, I just haven't experimented enough with the
technique to know where. Cutting at a colder temperature, at least with
OCT, tends to fracture the OCT - it looks and sounds like I am cutting
through a block of ice. I have tried a sample of Cryo-Gel from Instrumedics
which seems to need a colder temperature for sectioning but, contrary to
it's intended purpose, it is more difficult to work with. So it goes. I'll
try playing with the blade angle some more (my trainer would have an
aneurysm if she hear me say that), but what is the chance that will prevent
tissue loss? This is great though; I would like to keep the sectioning
conversation going (along with about half a dozen other issues), as
searching though the Histonet achieves doesn't produce much helpful
information. Cryo-sectioning, proper use of a cryostat, and what sections
look like when you do things right/wrong, as well as how to remedy common
problems, seems to be one of those grey areas where we are at the whim of
whomever teaches us. When I have some more time, I will contact you and Jan
about customizing my application.
Ion
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